ABSTRACT
During digestive organogenesis, the primitive gut tube (PGT) undergoes dramatic elongation and forms a lumen lined by a single-layer of epithelium. In Xenopus, endoderm cells in the core of the PGT rearrange during gut elongation, but the morphogenetic mechanisms controlling their reorganization are undetermined. Here, we define the dynamic changes in endoderm cell shape, polarity, and tissue architecture that underlie Xenopus gut morphogenesis. Gut endoderm cells intercalate radially, between their anterior and posterior neighbors, transforming the nearly solid endoderm core into a single layer of epithelium while concomitantly eliciting "radially convergent" extension within the gut walls. Inhibition of Rho/ROCK/Myosin II activity prevents endoderm rearrangements and consequently perturbs both gut elongation and digestive epithelial morphogenesis. Our results suggest that the cellular and molecular events driving tissue elongation in the PGT are mechanistically analogous to those that function during gastrulation, but occur within a novel cylindrical geometry to generate an epithelial-lined tube.
Subject(s)
Endoderm/embryology , Gastrula/embryology , Morphogenesis/genetics , Nonmuscle Myosin Type IIB/physiology , rho-Associated Kinases/physiology , rhoA GTP-Binding Protein/physiology , Animals , Animals, Genetically Modified , Body Patterning/genetics , Cell Polarity/genetics , Cell Shape/genetics , Embryo, Nonmammalian , Endoderm/cytology , Endoderm/metabolism , Gastrointestinal Diseases/congenital , Gastrointestinal Diseases/embryology , Gastrointestinal Tract/abnormalities , Gastrointestinal Tract/embryology , Gastrula/metabolism , Models, Biological , Nonmuscle Myosin Type IIB/genetics , Nonmuscle Myosin Type IIB/metabolism , Signal Transduction/genetics , Xenopus/embryology , Xenopus/genetics , Xenopus/metabolism , rho-Associated Kinases/genetics , rho-Associated Kinases/metabolism , rhoA GTP-Binding Protein/genetics , rhoA GTP-Binding Protein/metabolismABSTRACT
Early life stages of fishes have been shown to be especially susceptible to the toxic effects of heavy metal pollution. In this study, fathead minnow (Pimephales promelas) embryos were exposed in the laboratory to a graded series of aqueous methyl mercury concentrations under continuous-flow conditions. A number of toxicological endpoints were examined including; acute toxicity, bioaccumulation, protein production, impact on mitosis, gross and histopathology. Acute toxicity, reported as LC50 values of methyl mercury, ranged from 221 microg/l (95% C.I. 246-196 microg/l) for 24-h tests to 39 microg/l (95% C.I. 54-24 microg/l) for 96-h exposures. Fathead minnow embryos were shown to rapidly take up mercury from the surrounding water. Mercury levels in embryos reached levels of 2.80 microg/g wet weight after 96 h exposure to 40 microg/l methyl mercury. An initial elevation of total protein in embryo was observed in embryos exposed to 25 microg/l methyl mercury during the first 12 h of development. At later stages, significantly lower levels of protein/microg embryo were observed. Methyl mercury had no effect on mitotic stages (p=0.05) in early, cleaving blastula-stage embryos. Live embryos and serial sections were utilized to characterize changes in embryo morphology and histopathology.
